iNFF14

General#

Cell Line

hPSCreg Name RCSIi004-A
Alternative name(s)
iNFF14
Cell line type Human induced pluripotent stem cell (hiPSC)
Last update 11th October 2018
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Provider

Generator Royal College of Surgeons in Ireland (RCSI)

External Databases

BioSamples SAMEA5126964

General Information

Publications
Is the cell line available in principle? Available with restrictions: Available for non-commercial research

Donor Information#

General Donor Information

Sex female

Phenotype and Disease related information (Donor)

Diseases No disease was diagnosed.

Karyotyping (Donor)

Has the donor karyotype been analysed?
Unkown

Other Genotyping (Donor)

Is there genome-wide genotyping or functional data available?
No

External Databases (Donor)

BioSamples SAMEA5126965

Ethics#

We are currently revising this section.

hIPSC Derivation#

General

Source cell type
fibroblast of dermis
Source cell type (free text) Collected from foot
Passage number reprogrammed 5

Reprogramming method

Vector type Non-integrating
Vector Sendai virus
Genes
Is reprogramming vector detectable?
No
Methods used
RT-PCR
Files and images showing reprogramming vector expressed or silenced

Vector free reprogramming

Type of used vector free reprogramming factor(s)
None

Other

Selection criteria for clones To characterize colonies generated from SeV-infected skin fibroblasts, we picked an average of 20–30 colonies from each cell line. We utilized a live staining approach (Lowry et al., 2008) to select only the colonies that stained positively for both SSEA-4 (Millipore) and TRA-1-81 (BD Biosciences) antibodies confirming that the iPSCs were fully reprogrammed.
Derived under xeno-free conditions
No
Derived under GMP?
No
Available as clinical grade?
No

Culture Conditions#

Surface coating Non coated
Feeder cells gamma-irradiated mouse embryonic fibroblast
Passage method Mechanically
CO2 Concentration 5 %
Medium Other medium:
Base medium: DMEM/F12
Main protein source: Knock-out serum replacement
Serum concentration: 20 %
Supplements
Glutamax 1 %
b-mercaptoethanol 0.1 mM
NEAA 1 %
FGF-2 8 ng/ml
Has Rock inhibitor (Y27632) been used at passage previously with this cell line?
Unkown
Has Rock inhibitor (Y27632) been used at cryo previously with this cell line?
Unkown
Has Rock inhibitor (Y27632) been used at thaw previously with this cell line?
Unkown

Characterisation#

Analysis of Undifferentiated Cells
Marker Expressed Immunostaining RT-PCR FACS Enzymatic Assay Expression Profiles
SSEA-4
Yes
TRA 1-81
Yes
Oct4
Yes
Marker Present Absent
mCpG X
OCT4 X
Morphology pictures
Embryoid body formation; in vivo teratoma
Differentiation Potency
Endoderm
Ont Id: UBERON_0000925
In vivo teratoma
In vitro spontaneous differentiation
Marker Expressed
a-fetoprotein
Yes
Mesoderm
Ont Id: UBERON_0000926
In vivo teratoma
In vitro spontaneous differentiation
Marker Expressed
Vimentin
Yes
Ectoderm
Ont Id: UBERON_0000924
In vivo teratoma
In vitro spontaneous differentiation
Marker Expressed
BIII-tubulin
Yes

Genotyping#

Karyotyping (Cell Line)

Has the cell line karyotype been analysed?
Yes
46XX
Karyotyping method: G-Banding

Other Genotyping (Cell Line)